mericon Quant Legionella spp Kit

For real-time PCR detection and quantification of Legionella species in water

Highly sensitive and specific detection of Legionella DNA

Streamlined detection protocol

PCR run time of 73 minutes on the QIAGEN Rotor-Gene Q

Full license for PCR without additional costs

The mericon Quant Legionella spp Kit is designed for the target-specific detection and quantification of various Legionella species DNA in water after concentration (e.g., by filtration). The assay uses a real-time PCR-based protocol and forms part of the comprehensive QIAGEN food testing portfolio, which also features dedicated sample preparation kits and assays for ingredient authentication and GMO DNA detection. The mericon Quant Legionella assay performs optimally on the Rotor-Gene Q, but has also been validated for block thermal cyclers.

The mericon Quant Legionella spp Kit is intended for molecular biology applications in food, water, animal feed, and pharmaceutical product testing. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Performance

The mericon Quant Legionella spp Kit is for the specific detection and quantification of several Legionella species in water after concentration (e.g., by filtration). The assay can detect as few as 10 copies of Legionella DNA in a reaction and it exhibits high specificity for pathogens of that genus. In an experiment using 2500 copies of the tested DNA, no cross-reactivity was observed with other pathogens (see the table "Results of mericon Quant Legionella spp Kit cross-reactivity study").

Results of mericon Quant Legionella spp Kit cross-reactivity study

Pathogen

Result

Pathogen

Result

Legionella pneumophila

+

Legionella erythra

+

Legionella adelaidensis

+

Legionella feeleii

+

Legionella impletisoli

+

Legionella jordanis

+

Legionella israeliensis

+

Legionella tucsonensis

+

Legionella worsleinsis

+

Legionella beliardensis

+

Legionella spiritensis

+

Legionella lansingensis

+

Legionella brunensis

+

Legionella drozanskii

+

Legionella geestiana

+

Legionella nautarum

+

Legionella santicruci

+

Legionella shakespeare

+

Salmonella abony

–

Bacillus cereus

–

Clostridium perfringens

–

Campylobacter jejuni

–

Escherichia coli

–

Yersinia enterocolitica

–

Staphylococcus aureus

–

Listeria monocytogenes

–

Shigella flexneri

–

Cronobacter sakazakii

–

Principle

The mericon Quant Legionella spp Kit is designed for the quantitative detection of several Legionella species in water after filtration. Legionella species are most often found in water of warm-water systems, spas, pools, or cooling towers and are spread via aerosols.

Detection of pathogens using real-time PCR is based on the amplification of a specific region of the relevant pathogen genome. The amplified product is detected using target-specific fluorescent probes that bind to it. As the PCR product accumulates, there is an increased fluorescent signal from the bound probes. Monitoring the fluorescence intensities during the PCR run (i.e., in real time) allows the detection of the accumulating PCR product without having to re-open the reaction tubes afterward.

Quantification of the target Legionella species is carried out based on a standard curve covering a concentration range of 25,000–25 copies per PCR reaction. The standard curve is prepared from purified Legionella Standard DNA, which is provided in the mericon Quant Legionella spp. Kit at an accurately defined concentration.

mericon PCR Assays include an internal control to control potential inhibition. This aids in the straightforward interpretation of the results (see the table "Summary of possible PCR outcomes").

The PCR primer set for each assay is highly specific and targets a unique and conserved region of the tested pathogen genome that has been bioinformatically and experimentally verified.

Summary of possible PCR outcomes

Internal control amplified

Tested DNA amplified

Meaning

+

+

Sample is positive for the target DNA

+

–

Sample is negative for the target DNA

–

–

PCR failed due to inhibition

Procedure

DNA is extracted from filtered or concentrated water sample using a dedicated sample preparation kit.

The isolated, pure bacterial DNA is assayed using a streamlined real-time PCR protocol that returns results in 73 minutes on QIAGEN's Rotor-Gene Q. The protocol has also been validated for use on other thermal block cyclers.

The results are displayed as easy-to-interpret PCR curves.

Applications

The mericon Quant Legionella spp Kit is for real-time PCR-based detection and quantification of Legionella species in water.